Unidirectional and stage-dependent roles of Notch1 in Wnt-responsive Lgr5⁺ cells during mouse inner ear development

Wnt and Notch signaling play crucial roles in the determination of the prosensory domain and in the differentiation of hair cells (HCs) and supporting cells during mouse inner ear development; however, the relationship between the two signaling pathways in the mouse cochlea remains largely unknown. Here, we investigated the interactions between Notch and Wnt signaling on the basis of the bidirectional regulation of Notch1 specifically in Wnt-responsive Lgr5⁺ progenitors during different cochlear development stages. We found that the downregulation of Notch1 in Lgr5⁺ cells from embryonic day (E) 14.5 to E18.5 can drive the quiescent Lgr5⁺ cells to re-enter the cell cycle and differentiate into extra HCs, whereas the upregulation of Notch1 expression did not affect the proliferation or differentiation of otic progenitor cells. No effect was observed on the upregulation or downregulation of Notch1 in Lgr5⁺ cells from E10.5 to E14.5. We concluded that the roles of Notch1 in Wnt-responsive Lgr5⁺ cells are unidirectional and stage dependent and Notch1 serves as a negative regulator for Lgr5⁺ progenitor activation during cochlear differentiation. Our findings improved the understanding of the interactions between Notch and Wnt signaling in cochlear development.